CA2113268C

CA2113268C - Measuring molecular change in the ocular lens

Info

Publication number: CA2113268C
Application number: CA002113268A
Authority: CA
Inventors: Mark A. Samuels; Scott W. Patterson; Jonathan A. Eppstein; Nai Teug Yu; Sven-Erik Bursell
Original assignee: Georgia Tech Research Corp
Current assignee: Georgia Tech Research Corp
Priority date: 1991-07-17
Filing date: 1992-07-16
Publication date: 1999-01-12
Anticipated expiration: 2012-07-16
Also published as: US5203328A; ATE158704T1; EP0597932A1; ES2110007T3; EP0597932B1; EP0597932A4; JPH07500030A; AU661026B2; US5582168A; DE69222535T2; DE69222535D1; CA2113268A1; AU2373392A; WO1993001745A1

Abstract

Apparatus and methods for noninvasively diagnosing diabetes mellitus, the prediabetic condition, cataracts, and other diseases in the human (or other) body are disclosed.
Diagnoses are made by illuminating ocular lens (L) or other tissue with a narrowband light source (15) at a selected wavelength, separating the backscattered radiation into fluorescent and Rayleigh components (55, 135), detecting (85) the backscattered radiation intensity at the peak of the fluorescent response, and normalizing the detected value with the detected intensity of its Rayleigh component. Measurements provided in this manner can be used as improved indicators of the presence or absence of certain diseases or conditions.

Description

W 0 93/01745 -1 2~ 8 P ~ /US92/05941 MEASURING MOLECULAR CHANGE IN THE OCULAR LENS

This invention relates to ev~lu~ting ch~anges in biological tissues and more sI~eçifir~lly to d~a,a~s and mpthods for ;vely mP~cllring molecular c~n~eS in the lens of the eye.

R-r~.ound of the Il~,cn~ol-PY;Ct;~ odc for d;a&,~r:n~ A;~S, par~cuLarly ,l;~t~ s, are often less ~an desirable. One such mPth~d, ~e o~
~hlcosp~ tolerance test, ~Ih ..~ c to a sist 11ia~nocic of r~ ch~s mP.llitlls by d~ r e1: ~,a~ blood gl~,co~ levels exist in ~ ;. nlc sll~.t~l of having the disease. ~use many p~ ~ having 10 cle~lat~ levels fail subsequently to de~elol, the c~ r~ t~,...c of the disEase, however, the re~abi~ly of the test is grn~ ally qllPS~ion~.
A second Aia~n~stic m~!hod~ ~e Islet Cell Alltibody aCA) test, may be used to predict those paticnts at risk for type I Ai~hetes and can pl~te the onset of de~ilits ing c1i~ 1 sy.-~ -..c by as much as five years. The ICA test is not qpically lltili7~d, h~ , be~1se of its complexity, e~ ~, and lack of specifi~ity and bccause of a lack of ~ l;r n among evsl~ng l~\}JO ~ ;f'S. rull~ G~e, the test is useful only for ~1etecti~ type I diabetes, which strikes only ap~ro~ ;-n~t-;ly ten ~rwl ~ of ~e entire Ai ~eti~ patient poF~ tion. By cQntr~ct ~y~ t~ of having the ~A~I;r co-~A;~ for ~ypê
II diabetes c;u~ ly have no cQ-~r.~.r --8 diagnostic procedure.
It is well known ~at cer~in ~.l;ons of the eye fluoresce when i~ At~. The lens of the eye, for eY~rnr'A, can be made to fluoresce intense1y when i1~ ~ wi~ radia~ion having a waveleng~ bet~en ap~,-o~ tP-ly 350 nm and 550 nm. Utili~ng rAAi~tion of a waveleng~ less ~an a~ u~ Ply 400 nm typically is avoided ~unless power levels and e~ mes are restnc-ted), WO 93/01745 ' PCr/US92/0~941 however, since this higher Çl~quency radiation is known to cause d-q-mage to ocular tissue.
The p~sence of certain ~ q-~es in the hurnan body cause chemir~l chq-nges in the lens of the eye, q-l~ering the mol-nt of the S fluolesccnt ~ onsc to an ;11'J''';~ ;OI~ of thc lens. The lenses of c~t~acl p~ nl~, for example, be~ opaque due to lipid :)~;d~;OI, protein ~ co~ Lon, and the co~ sion of sullLydl~l (-SH) bonds to ~ lfi~e bonds (-SS). Similqrly~ in ~liq~etps mPlli~l$ and g,q~ ose-..;q, the gl~lco~ and ~,q-l~ctose are converted to sorbitol and 10 dulc;~l, lcspc~ ely. Ac~mlllqtio~ of these c~,ll~ounds results in a high osmotic g~iPnt within the lentirlllqr cells. Prolong~ Illc with dn~gs such as cor~costeroids and chlo~ also causes o~r~iti~s of the hunun lens.

U.S. Patent Nos. 4,895,159 and 4,883,351 to Wciss thus, ~ Qse meth~d5 for ~t~ g ~e cxistence of ~iq~e~es using light scat~l~d from l~nticlllqr cells. As ~esc- ;he~ in the Weiss patenLc, the ba~L~ led light from a patient S~ of ha~ing ~i~bet~ is used to c?l~.llqtJe a fliffilcion c~P-ffiri~nt for that pti~nt A
20 second dct~ l;Q~ of diffil~ n coPffi~ -ntc iS made for a control group of nl~n~iqhe~ic p~ t~, and the ~ ;on cG~ l of the s~lspectP~l rliqhetic is co~ d with those of r~o~di~betic, control group ~I;I~n~c of the same age.
Re~quse lenses typically cloud n~tllrqlly as p;~tientc age, 25 however, meas-lre.l~ made in conn~ction with the methods of the Weiss patents can be taken only from clear sites in the ~qtientc~
lenses. The Wciss ~h~ Jes also appear unable to ~licting~ h the IlltimqtP cause of cl~ng~ in diffiJsi~n coefficient~ or to detect the prediq~etic corldition (i.e. where no overt clinical signs of ~~iqhptes are wo 93/,~l74~ Pcr/us92/oss displayed but will be exhibited ~,vithin ap~io~ ..qtPly five years as, for e~llplc, when a positive ICA test occurs), sinc_ myriad ~ q-oes and physiological con~itiQns are known to affect the lens in the ..,~nn~.r therein described. Use of the ~ ion co_fficient as a stand-alone S diagnostic test also suffers firom its vPri~ility as a function of patient age, par~cularly since results have both age~epe~e-nt and age-çnl ~".. ;z~rc .

Other patents, such as West Gernun Patent No. 261957Al to Schiller and U.S. Patent No. 4,852,987 to Lohmann, d~s~,;he P~ q~, diagnostic ...- ~I.ods in which the fluo~-~cc signaI
c;l;rs are CO~ . The S~ patent, for exampb, di~loses CQ-..~t;~ fluolesc~llce signal intenCities at two wavel~nE~lls using a single e~ t~tion wavelength in an cffort to detect the presence of 15 cqtq-r~q~ctc. The raho of the r~llting fluolcsc~--ce int~ es is c~ r~d to the ratio obtau~ed at the same wa~l~thc from known cataract ~I;- nlQ to achieve the desired ~1iagnostic result. As dcs~--;l~
in the Schiller patent, the exçit~ti~n wavelengths are s~lrct~ from the ranges 32~340 nm, 380-390 nm, and 43~450 nm, while the i~nc;(~
20 of fluoresc~P-n~e peaks is ~ t within ~.a~cl~nglh ranges of 410-440 nm, 450-460 n n, and 50~520 nm. In con~ct to the .CChillPr patent, the T4k-..z~ patent Illeas~,s thc n~gnitude of fluolosc4n~4 nc;l~ at a single wavelength created loy light of one eYcitq~irn wavelength and co",~s this ;~ Q 1,~ to known ~ n~;l;ÇS at ~c 25 given wavelengths in order to det~",line the degree of eye lens clo~l-liness. Neither of these patents, ho.._~er, teaches or s~ggectc detection of ~i~hetçs or the preAi~betic co~ n.

.~

wo 93/01745 Pcr/Us92/059 ~ 4 ~
3.~,3 .Uc~ of ~e Invention The l.~senl invention provides a~ nl~c and methods for nol~vasively ~liqgnosing se1~-ct~ i~q-~s, i~r1~ g .li~,~s and the preAi~'~etic condition and various diseases qrrP-c~ g metabolism, in S tissues of 1,--."~--.c or other ~qnimq1c. Utilizing a lu~u~v-band light source of w~ gth typically ~;h.~ l 4~1500 nm (and, p,~ f~ly, appi~o.;-..~P.1y 406.7 nm) from a laser or similar device and a confoc l lens system, the pl~se.ll invention ;11~J .~;n~,s the ocular lens (or other) tissue and d~ .es ~e intensity of ~e b~fiL~c~ d ~"li~ n at lO bo~ the peak of the nuO,~SCenl l~SpO~ (typically at ap~ 1y 490 nm within the range 460 1800 nm) and the peak of the Rayleigh cc-.~l~nF~ (at the eYr~-q-tinn ~.a~C~ ). The d~h ~h~ radiation uenlly is transmitted to a spectlo.~ r to be divided into its various cG,..~i~n~ (e.g. fluo~sc~ G and Rayleigh). The ;-~t~ y of 15 the flu~l~sf~nl co~ on~n~ is then n--~qli7e~ to the intensity of the Rayleigh cs..~l,u..f-~ by forming the ratio of the fluol~cscent intensity to the Rayleigh i~tcn~:ly. The ~ .oun~ of the ba~c~
fluol~sf~lll and Rayleigh r9q~1iqtinn p~ ide a reliable indi~qtor of the onset and ~rO~SSiOI of ~ ~,s such as (but not n~c~sc -. ;1y liTnited 20 to) ~ b~tcs me11ih1c, the p~ ;c co~itiQn~ and c~prs~rtc in the human or other body.
Unlike e-Yicting ~ ~hn ~eS such . s those desc~ il-cA above, the ple5el l invention essrnl;Ally e1;-..;na~-s the age~e~?endenl "-eas.ll~-ll~.l v-q-ri~ions previously shown to be ~l~senl. By ~n~q~uri-lg 25 the Rayleigh co~ on~nl of the b~ diqtinn and using it for normA1i7A~ion, the precise ~ Qv~ of i11~ ;nn energy delivered to the subject lens tissue area l~,lative to the q-n~r11nt of fluo,~sc~-nre signal ge-~e~ A by the tissue can be ~ct~ l. This approach reduces comp1irqtionc A~Cociqt~d with vq-r ~nres in lens opacity which wo 93/01745 2 1 i 3 ~ ~ 8 Pcr/US92/05941 can alter, in an un~no~v~ ollion, the level of ilh~ ;on delivered to the subject area. By ~ .ng the effect of the i,~;ecls' ages on the test results, the ~,rl ni~P pe-.~ es~bl;c~ r-~l of a clear ~reshold--d~ of ag~ ;~ the ~ heti~ and p~ ir, ~ "~j 5 from those wi~luul the disease. The il,~en~oQ also neither leq~li~S
use of a coh~n~ Iight source nor suffers from the hck of sperifirity (eYi~tinp in, e.g., the Weiss ~ch~ es) in ~;s~-;-..;-~l;-~g the ultim~
cause of the effect being measured.
It is ~ .,fole an object of the yl~ t invention to provide 10 a~ s and .~ ~s for non.~ iagrnsing ~ s mP11itl1c, the y~ el;c co~ ;l;o-~, cataracts, and the pl~3~nce of other ~ic-p~se in~h1rli~ ones ?rf ~ g met~abolism.
It is ~no~l.rr object of the ~ t invention to ~.~vide uc and ..-~1.~s ~-... Il;~g nr~mUli7 ~ion of a fluo~sc~-nce 15 signal scd~led from a subject eye or other tissue by the Rayleigh c~n~l onrn~ of the sc,.~ d ,~--1;- 1;....
It is a further object of the p~nl invention to provide app~ua~s and mPtho.1c es~-~l;qlly e1;-~ g the age-depc-.de-.l ~I-eas.~ nl v? iqtinns previously shown to be ~ nl in eyicting 20 ~iqgnnstiC h-ch-~ Gs.
It is yet ~qnothPr object of the p~ nl invention to provide al ~lus and metho~l~ for .~ g the lens tissue over time for, e.g., cv~ qtin~ the efficacy of ~ s m~llihlc l,~l.... ~.l or pl~enl~ e l~chniqve~ deqlin~ with the ~.~A;A1~!;C condition.
~ 25 Other objects, r~lu,~s, and adv-q-n~ges of ~e pl~senl invention will become appalenl with lefel~. ce to the le-~;nder of the written portion and the drawings of this ~li~tion wo 93/0174s 8 pcr/uss2/o5941~ 3~,6 ~

Brief Des.i,i~lion of the Drawin~
FIG. 1 is a srl-f~ es~ ;on of an a~ s of the p~ nl invention.
FIG. 2 is a sc~ n~-~;on of an ~lI. ,"~t S e.l.bo l; f-nl of the ~f.~ s of F~G. 1.
FIG. 3 is a g~aphic~ I;nn of the fluo~sce.lt signal intensity as a r~ n of age of both diabetic and nondiabetic p~tientC ob~illed using the ~ ~s of FIG. 1 as ~es~ ~l in the EXAMPLE herein.
FIG. 4 is a ~phie~ t~l;r)n of the ratio of the fluo ~sc~nl to Rayleigh signal intensities as a r. ~c1;o~ of age of both diabetic and nondiabetic patients ol~ ~ using the apparatus of FIG.
1 as des~-~ibe~ in the EXAMPLE herein.
F~G. 5 is a gl~ph;~ he~ ;~n of the fluo~s~ll 15 signal; ~.t ~c;ly as a r..-~ of age of both diabetic and ~o ~A;~l~e1ic ob~in~ using the ~ .c of FIG. 1 for an m ....;~ n ~iq1ion wavrle ~gll. outside a p.ef~.lcd range of that used in connection with the present l,l~e.lLon.
FIG. 6 is a ~rhi(ql l~ ;nn of the ratio of the fluolescel t to Rayleigh signal int~.~lies as a filn~tinn of age of both eti~ and n- n~ ;G patients obt~ned using the ~ a~lvc of FIG.
1 for an ill~----;n~t;~n rvq~liq~ n ~.a~relen~ outside a p~L Qd range of that used in connection with the ~l~sen~ iol,.

Detailed Des~;~ion FIG. 1 illustrates an optical system 5 of the pl~s~nl invention. Optical system 5 inc~ es a light source 15, lens 25, a COI~Cdl lens system 35, collector 45, and a spe~ ~r 55. Source 15, which provides l~ru~-band ii1...~ u~;nn, typically may be a low -wo 93/01745 Pcr/uss2/os941 w 7 ~113~68 power k~ n laser tuned to produce r~ 1iqtinn having a wavelength ~ bel~n a~pro~ 1y 400-l500 nm. In one embof~;.. ~nt of optic~
system 5, source lS plo~ides r: ~iqti~ n at a wave1~n~th of 406.7 nm.
Also shown in FIG. 1 are ocuL~r lens tissue L, 5~ "~to~ 65, eyepiece 5 75, d~t c~ and l,loces~ assembly 85, and fiber optic ~ .,guides 95 and lO5.
Accol.Lng to FIG. l, ~ -.u~t~r 65, used to reduce the power level of the ~ c...~ radiation, ~ es ~iq~on from source 15 and f~ ds it to lens 25. Lens 25, which may be a 40X
10 miclosco~ objec1ive or other similar device, then f~u~s the (zl~ adiation onto ~e end of ~a~uide 95, which in turn l~r~c~ the ~iqticn to con~ocal lens system 35. Lens system 35 s.ll,s~l.,en1ly deL~.els the ,~ ;of to a ~1e.~ ~r~?-~ nP of ocuLar lens tissue L (typically apl.r~ y 200 cubic mi~ i). A mr~1i 15 slit lamp base may be used to house and po";Linl~ lens system 35 for easy access to lens tissue L, while lens system 3~ itself is ~P-si~rled to permit the same ~olu.-.e of len. tissue L to be held in the focal point of col1Pctor 45. In an e-~h~~ fnl of the pl~ t invention con-:-t~
with FIG. 1, the ~l)c,t~r~ 115 of lens system 35 at its focus is greater 20 than ap~loA;~ y fifteen miclo...~tc-~, &,ic...;,~ that the excitqtion r~iqtirn di~ges rapidly after p~c~ g through the focal point of lens system 35 and IL~ c;~ ~e ~pot intensity of ~e ~
should it e~-ro~ any other portions of the ocular tissue.
Collector 45 receives the rtq-liqltion b-q-r~c~ d from lens 25 (or other) tissue L as a result of it being ill~l.ninqvtPA by rq~liqticn fromsource 15. From collP-ctor 45, the bacL~ d r~iqti~n is directed into wav~g~lide 105 and ~.a...c....ll~d to ~e entrance slit 125 of the mr~nr~hro",alor 135 forming spe.;~u -,~. 55. If desired, col1ectQr 45 also may direct a portion of the b:~r1~Sc~ tirn to eyepiece 75, WO 93/01745 PCI'/US92/05941 30 ~ 3 -8-p ~ g an ope,ator to view the exact location of the S~
volume of lens tissue L.
Division nd p,ocessing of the bY~ ~d ~diqti~n occurs in s~cll~o",~t~r 55 and d~t~-c~ n and pn)c~sC;~g !qc~semhly 85.
S Radiation l~n~ d to s~cllo"~ 55 initially is .~p~ f~d into its Rayleigh and flolc~ e C~ f~ . The two co--~~ubs~ e~ y are directed, l~spe.~i~ely and as neC~s~y~ to ~ e fo-.-.;~ part of assembly 85, for d~ t;on of the intensities of each. Assembly 85 also may i~ de a digital computer or similar 10 computing device for fs .--; ~g ~e ratio of the fluo.~ seelll and Rayleigh co..~l~nr~ of the ba~ iqtion, l~ nor~qli7i~ the peak intensity of the fluon~s~.~l co--~ -n~
An ?lt~P'~qtP,~ e~-~ho~ --f -~ 10 of optical system 5 is inlls~tp~ in FIG. 2. Acco~ g to FIG. 2, light source 20, which 15 may be a laser diode, produces radiation of ~ t~ ap~lo~;"~ y 813.4 nm (wi~in the range of approxinutely 800-860 nm) and is coupled to a no~ )f~r L~u~n~ d~!~V~hli~ device 30 to produce the desired ~.a~ g~ output of 406.7 nm (within the range 40~1500 nm). Light source 20 v~ y may be a laser, light emi~
20 diode, or other n. rrow-band light source (jnrl~ o~ &ouices co ~ to optical filters). The lqd~lJ;on ~ sc~ Pntly is directed through an optical delivery ~ystem 40 into the eye 50 of a p~Pnt As with the optical system S of FIG. 1, -q~ c,ll'~o~ nl 10 i n~u~les an optical cQllectQr 60 confucal to the del~e.~ system 40 to collect the 25 b~ c~ d r;~iqtirn from the lens of eye 50. Simil-q-rly as noted above, the bac'i~c~ iqti-n conected in~ les both a . fluo.~scP~-c~. sign l (typic lly appro~ P-Iy 490-500 nm within the range 460-1800 nm) and an intense Rayleigh co..-~o~ at the ill.. ;n~;on wavelp-r~th-WO 93/01 745 211 ~ 2 ~ ~ PCr/US92/0~941 _9 FIG. 2 ~ ldition~lly di~loses means for sC~n~ g the COIllyOllflllS of interest of the ba~calf~ radiation, ;"~
dichroic beam S~ S 70 and 90, and for d~-f~-c~ g the ;nf~n~ of the CGIll~ll~,.ll~ c;~mllPneo~ls1y using single chip silicon d~lu.~ 100 and 5 120 or similar devices. All ~. ~f;./ely, co~ onfn~ ~s~ ;nn may be ~~col.~ hf~ using beam ;,pli~.~ in Co~ ne!;~n with opticall,~.d~c~
filters or d;~ e c~ r!~t~ such as ~1;rr.~cl;~ n g-~ . Hybrid d~ t~lor/filfer assemblies also IlUy be used. Elec~onic ci,.;uil,~ 130, such as but neither limited to nor n~c~s~-.;ly ,~.~;.;..g analog 10 z-.~l l;r.f -~, analog to digital (A/~) C~ , and a digi~l co..~
yroc~s~s the data det~chd by ~lcb~c~ 100 and 120, c-q~ qt~-s the nnrmqli~d fluorc~.l~yleigh c~ e ~ ratio, and, if desired, makes the result available to an operator through a digital display or other sllit~'~le means. Eyepiece 80, finally, may be used by ehe 15 upe-~lo~ to view the hcqti~n of the cytitq~n focal point in eye 50.
The ples~ iol~ may further be ulld~k)od wieh ~Çelence to ehe following non~ g EXAMPLE.
EXAMPLE
FIGS. 3-6 ilh~strq-t~ data ob~;l ed from rli~ -ql trials 20 con~luct~l using sixty-nine (69) human y~ i aged twelve (12) to sixty-five (65). Forty-eight (48) of ehe y,~ c had previously been di-a~r~o~ as having diabetes, while the ~ g twenty-one (21) had not. F~GS. 3 shows ehe total fluo~ --ce signal ob~ e~ for each yatient (e~y~ssed in "Counts x 105," where the ~.. l~r of Counts is a 25 fiilnrti~nn of the ,.~ er of emitt~d photons per unit time) using an ill...~,;.~ljnn wavelongth of 406.7 nm. FIG. 4 details the results when those same fluorescenre signals are norrnqli7ed by the Rayleigh co-"~ne,-l of the ~acL ~r~lt~ tinn in accor~ ce with the yl~nl invention. As ill..~ ~ in FIG. 4, ,q,lth~-gh the norm-q-li wo 93/0174~ Pcr/uss2/o594l 3~

signals trend u~w~ud as a filrlG~ion of age, they evidence clear ~;C~ r~;ons bet~.~n those p~l;f~ known to have ~ ~s or the ;~ con~ ;on nd those who did not. The nonnqli7~d signals for the nOn~ S, for example, were less than lI,i~Rn (13), while S those for diabetics excee~e~ fifteen (15).
By cc-~ , use of an ill---.-;--;-l;rn wa~ e-.,eth of 441.6 nrn (outside the range of the ~l ~enl ,.,~e.llion) produced less desirable results. I;IGS. 5-6, which cV-~ pQn~ spe~ ely~ to FIGS. 34, show (in FIG. 6) less of a distinc~on ~h.~n the nnn~Lqli7~d signals 10 for the ~liq~etic as o~ to .~on~ ;c patients. I~u~lh~--...~re, those p~l;~--l'~ who tested ICA positive are shown to have flucl.,scen JRayleigh ratios within the range of nondiabetic patient values. As a result, no clearly established threshold is available for ~i~q~nos1ic ~ Jo~S.
The rolegolng is pi~,.ided for ~sses of i~ ,-l;n--, exphnation, and ~e~~ ;nn of e-~ o~ e-~ of the present invention.
Morlifiratinnc and ;vl~p1-l;nnc to these en~l~v~ ellts will be a~
to those of O~il~ skill in the art and may be nude wi~oul depal~ulg from the scope or spirit of the i,l~.lliol .

Claims

We claim:

1. An apparatus for measuring molecular changes in a patient having an ocular lens that, when illuminated, is capable of backscattering radiation including fluorescent and Rayleigh components of determinable intensities, comprising:
a. means for illuminating the ocular lens with light having a selected wavelength, thereby causing the ocular lens to backscatter radiation in response to the illumination;
b. means, responsive to the backscattered radiation, for collecting the backscattered radiation;
c. means, connected to the collecting means, for separating the backscattered radiation into its fluorescent and Rayleigh components; and d. means, connected to the separating means, for (i) detecting the intensity of each of the separated fluorescent and Rayleigh components and (ii) normalizing the detected intensity of the fluorescent component by the detected intensity of the Rayleigh component, thereby producing a measurement of molecular changes in the ocular lens.

2. An apparatus according to claim 1 in which the illuminating means comprises:
a. a light source selected from the group consisting of lasers, laser diodes coupled to nonlinear frequency doubling devices, light emitting diodes, and broadband sources coupled to optical filters;
b. a lens, optically responsive to the light from the light source, for focusing the light; and c. a lens system, optically responsive to the focused light, having a focus, and defining an aperture at its focus greater than approximately fifteen micrometers.

3. An apparatus according to claim 2 further comprising an eyepiece, responsive to the backscattered radiation, for permitting an operator to view the ocular lens.

4. An apparatus according to claim 3 in which the separating means comprises at least one dichroic beam splitter.

5. An apparatus according to claim 4 in which the detecting and forming means comprises at least one single chip silicon detector and the wavelength of the fluorescent component of the backscattered radiation is between approximately 460-1500 nm.

6. An apparatus according to claim 5 in which the detecting and normalizing means further comprises an amplifier.

7. An apparatus according to claim 6 in which the illuminating means further comprises means for adjusting the power level of the illuminating light.

8. An apparatus for measuring molecular changes in a patient having an ocular lens having a volume that, when illuminated, is capable of backscattering radiation including fluorescent and Rayleigh components of determinable intensities, comprising:
a. a laser for providing light having a selected wavelength and power level;
b. means, optically responsive to the provided light, for adjusting the power level of the light;
c. a lens, optically connected to the adjusting means, for focusing the light;
d. a first optical fiber, optically connected to the lens, for receiving the focused light;
e. a lens system, optically connected to the first optical fiber and defining an aperture having a focus greater than approximately fifteen micrometers, for delivering the focused light to a selected approximately two hundred micrometers of the volume of the ocular lens, thereby causing the ocular lens to backscatter radiation in response to the delivered light;

f. a collector (i) having a focal point encompassing the selected volume of the ocular lens to which the focused light is delivered and (ii) responsive to the backscattered radiation, for collecting the backscattered radiation;
g. a second optical fiber, optically connected to the collector, for receiving the collected radiation;
h. means, connected to the collecting means, for separating the backscattered radiation into its fluorescent and Rayleigh components; and i. means, connected to the separating means, for (i) detecting the intensity of each of the separated fluorescent and Rayleigh components and (ii) normalizing the detected intensity of the fluorescent component by the detected intensity of the Rayleigh component, thereby producing a measurement of molecular changes in the ocular lens.

9. An apparatus according to claim 10 in which the wavelength is approximately 406.7 nm, the separating means is a spectrometer, and the detecting and normalizing means comprises a computer.

10. An apparatus according to claim 8 in which the separating means comprises at least one dichroic beam splitter.

11. An apparatus according to claim 10 further comprising an eyepiece responsive to the backscattered radiation, for permitting an operator to view the selected volume of the ocular lens, and in which the measured molecular changes assist in diagnosing conditions selected from the group consisting of diabetes, the prediabetic condition, and cataracts.

12. A method for measuring molecular changes in a patient having tissue that, when illuminated, is capable of backscattering radiation including fluorescent and Rayleigh components of determinable intensities, comprising the steps of:
a. illuminating the tissue with light having a selected wavelength, thereby causing the tissue to backscatter radiation in response to the illumination;
b. separating the backscattered radiation into its fluorescent and Rayleigh components;
c. detecting the intensity of each of the separated fluorescent and Rayleigh components; and d. normalizing the detected intensity of the fluorescent component by the detected intensity of the Rayleigh component, thereby producing a measurement of said molecular changes.

13. A method according to claim 12 in which the illuminating step comprises the steps of:
a. providing a light source selected from the group consisting of lasers, laser diodes coupled to nonlinear frequency doubling devices, light emitting diodes, and broadband .
sources coupled to optical filters, for emitting the light at a wavelength of approximately 406.7 nm; and b. focusing the light using a lens system having a focus and defining an aperture at its focus greater than approximately fifteen micrometers; and further comprising the step of comparing the ratio of the detected intensities against at least one preselected value for assisting in diagnosing conditions selected from the group consisting of diabetes and the prediabetic condition.

14. A method according to claim 13 in which the detecting step comprises the step of detecting the separated fluorescent component at a wavelength between approximately 460-1800 nm and in which the comparing step comprises the step of comparing the ratio of the detected intensities against two preselected values, thirteen and fifteen, so that if the ratio is less than thirteen the patient may be diagnosed as unlikely to have the selected condition and if the ratio is greater than fifteen the patient may be diagnosed as likely to have the selected condition.

15. A method according to claim 14 in which the separating step comprises the step of separating the backscattered radiation using at least one dichroic beam splitter.